Hemoglobin ELISA Kit - bei Immundiagnostik

Arbeitsanleitung / Manual human Hemoglobin
1. INTENDED USE
The Immundiagnostik Assay is intended for the quantitative determination of human Hemoglobin
in stool. For In-vitro_diagnostic use only.
2. INTRODUCTION
In contrast to other commercially available Hemoglobin quick tests, this Hemoglobin ELISA
does not require previous adherence to a diet (no raw meat etc.) and recognizes human Hemoglobin
in 100-fold lower concentrations. This avoids false-negative results. Because of the
choice of antibodies, false-positive results are almost excluded.
Recent data show, that the clinical specificity and sensitivity can be increased by using Hemoglobin
ELISA.
Indications
· Occult blood in stool
· Crohn´s disease; Ulcerative Colitis
· Suspicion of colon carcinoma
· Polyps in the colorectum
3. PRINCIPLE OF THE TEST
The Enzyme-Linked-Immuno-Sorbent-Assay (ELISA) is used for quantitative determination
of Hemoglobin in stool. The Hemoglobin in the sample is bound to anti-Hemoglobin antibodies
(in excess), which are immobilized on the surface of the microtiter wells. To remove
all unbound substances, a washing step is carried out. In a second incubation step an anti-
Hemoglobin peroxidase labeled antibody is added. After another washing step, to remove all
unbound substances, the solid phase is incubated with the substrate, tetramethylbenzidine
(TMB). An acidic solution is then added to stop the reaction. The color converts from blue to
yellow. The intensity of the yellow color is directly proportional to the concentration of Hemoglobin
in the sample. Hemoglobin, present in the patient samples, is quantified by referring
the optical density of patient‘s samples to a Lot-dependant master calibration curve. This is
done by using Reference/Calibrator that is run with each test.
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